Table 3.

Effect of point mutation of EpRE sequence on EpRE activity

EpRE	Sequence	Constitutive activity (mean ± SD)	Induction fold (mean ± SD)
EpRE4	5′-CCTCCCCGTGACTCAGCGCTTT-3′ (wild)	2.16 ± 0.20*	2.47 ± 0.36*
	5′-CCTCCCCtTGACTCAGCGCTTT-3′	1.64 ± 0.27*	1.16 ± 0.24
	5′-CCTCCCtGTGACTCAGCGCTTT-3′	2.05 ± 0.31*	2.31 ± 0.48*
	5′-CCTCCtCGTGACTCAGCGCTTT-3′	2.21 ± 0.18*	2.15 ± 0.26*
	5′-CCTCtCCGTGACTCAGCGCTTT-3′	1.94 ± 0.28*	2.29 ± 0.17*
	5′-CCTtCCCGTGACTCAGCGCTTT-3′	1.49 ± 0.12*	1.15 ± 0.22
	5′-CCcCCCCGTGACTCAGCGCTTT-3′	1.46 ± 0.13*	1.08 ± 0.14
	5′-CCTCCCCGTGAaTCAGCGCTTT-3′	1.15 ± 0.24	0.86 ± 0.21
	5′-CCTCCCCGTGACcCAGCGCTTT-3′	1.96 ± 0.19*	2.13 ± 0.18*
	5′-CCTCCCCGTGACTtAGCGCTTT-3′	1.06 ± 0.11	1.17 ± 0.28
	5′-CCTCCCCGTGACTCAatGCTTT-3′	1.47 ± 0.27*	1.08 ± 0.19
	5′-CCTCCCCGTGACTCAGCtCTTT-3′	1.71 ± 0.25*	1.32 ± 0.29
	5′-CCTCCCCGTGACTCAGCGtTTT-3′	2.09 ± 0.21*	2.38 ± 0.35*
	5′-CCTCCCCGTGACTCAGCGCccc-3′	1.45 ± 0.17*	1.22 ± 0.31
n8	5′-GTTTCCCTTGAGTCAGCGCGTT-3′(wild)	1.77 ± 0.21*	0.82 ± 0.33
	5′-GTTTCCCTTGAGTCAatGCGTT-3′	1.87 ± 0.25*	0.97 ± 0.21
	5′-GTTCCCCGTGAGTCAGCGCtTT-3′	1.96 ± 0.28*	2.37 ± 0.34*
EpRE3	5′-CCGCCACGTGACTGCGCGGGCC-3′ (wild)	1.51 ± 0.27*	0.87 ± 0.24
	5′-CCGCCACGTGACTGCatGGGCC-3′	1.69 ± 0.27*	1.13 ± 0.18
	5′-CCTCCACGTGACTGCGCGcttt-3′	2.03 ± 0.22*	2.27 ± 0.36*
n1	5′-TCAGTAAGTGATGGAGCCTAGA-3′ (wild)	0.96 ± 0.13	0.88 ± 0.31
	5′-TCAGTAAGTGActcagcCTAGA-3′	1.58 ± 0.27*	1.26 ± 0.25
	5′-CCTCTAAGTGATGGAGCGTttt-3′	1.12 ± 0.17	0.97 ± 0.17
	5′-CCtcTAAGTGActcaGCgTttt-3′	1.93 ± 0.24*	2.35± 0.46*

Note: the constitutive activity is referenced to vehicle vector (pGL3 luciferase promoter vector). Induction fold is the division of luciferase activity of HNE exposure and that of vehicle exposure (ethanol). Putative core EpREs were underlined and mutated nucleotide was in italic lower case. N=3,

^*P<0.05 compared with vehicle vector (Constitutive activity section) or without HNE exposure (Induction section).